Complicated vital activities often result from synergism of a group of biomolecules serving as core execution elements. Fingerprints of many diseases remained in a molecular level are often decided by multiple biomarkers. In other words, “pattern characteristics” presented by multiple marker sets have stronger correlation with specific physiological and pathological phenomena than those of a single molecule. It is estimated (Nature, 2011.469(7329):p. 156-157) that thousands of biomarkers have been described in about 15 millions of documents, among them, there are about 100˜200 common markers approved to be clinically used. Relative to a single biomarker, a biomarker set consisting of multiple characteristic molecules is capable of providing crucial instruction information for clinical works in the aspects of early diagnosis, disease subtypes, risk predication, drug selection, disease course monitoring, prognosis estimation and the like in a more comprehensive, precise and efficient manner. In life science research, analysis on multiple biomarkers are often used for disclosing an action mechanism of a biological signal path. In addition, a large number of multiplex biomarker detection is introduced in modern pharmaceutical research for evaluation of drug efficacy and toxicity. As precise medicine age comes, precise quantification on multiple biomarkers in a large number of specimens will increasingly become an indispensable mainstream analysis task in the fields of modern diagnostics, therapeutics, drug research and development, translational medicine and the like.
A multiplex biological detection technology is a powerful tool satisfying a large-scale parallel biological analysis demand. Undoubtedly, although some classical single detection technologies (such as enzyme-linked immune assay, immunoblotting and real-time quantitative PCR) can also complete quantification on multiple analytes through a large amount of complicated operations, it is still difficult to overcome the following obstacles: respective sampling directed to different analytes leads huge logistic burden (sample collection, record, storage, transportation and the like); the same specimen must be allocated to multiple different detections, but its limited volume difficulty considers all the detections (especially oncology department and paediatric specimens); the whole set of detection is expensive in cost and huge in manpower consumption; timely completion of all detection items faces a huge pressure. With HPV (human papilloma virus) detection as an example, 14 high-risk type HPVs, affirmed by WHO (World Health Organization) and IRAC (International Agency for Research on Cancer), among about more than 30 HPVs that have been discovered at present and can infect human genital tract mucosa, can easily lead generation of cervical cancer. Considering necessary control and 3 repetitions, about 60 routine PCR (polymerase Chain Reaction) reactions need to be done for the whole detection, burdens of subsequent cost and operation amount make it difficulty popularized in clinic. Facing an increasing automated high-throughput multiplex assay demand in clinic diagnosis and medicine research and development application, a single detection mode of a routine technology has become a bottleneck restricting data output efficiency. However, for multiplex assay, all the detections can be completed in once operation without separating specimens, thereby not only greatly reducing the consumption of detection and shortening the detection period, but also facilitating standardization of sample treatment and automation of measurement methods.
A suspension chip is a technology utilizing a sorting coding microchip suspending in a liquid phase as a reaction and signal detection carrier. Its core characteristic lies in that the used microsphere has a code that is unique and can be identified by a machine. At present, the most widely applied suspension microchip technology mainly includes a fluorescence-coded microsphere technology and a bar code magnetic bead technology, but these suspension microchips generally have the deficiencies of complicated preparation process, high cost and the like.
The inventor of the present application also once developed a coding graph suspension chip based on a high-refection multi-layer dielectric stacking plated film, optical contrast between the code and a base mainly depends on a reflection effect of a high-reflection plated film. Under the illumination conditions of common white light, it is needed to perform film system design aiming at light intensity distribution of a specific light source, and it is need to perform high-precision plating process many times to achieve an expected reflection effect, so process difficulty is large and manufacturing cost is high.